The mAbs anti-HIV-1 p24-FITC (clone KC57), anti-Vα24-FITC (clone C15), and anti-Vβ11-PE (clone C21) were from Beckman Coulter, anti-CD1d-PE (clone CD1d42), anti-CD3 AF700 (clone UCHT1), anti-CD4 BV605 (clone RTA-T4), anti-CD11c-allophycocyanin (clone B-ly6), anti-CD11c PE-Cy5 (clone B-ly6), anti-CD45 PerCP (clone 2D1), anti-CD56 AF700 (clone B159), anti-CCR5 APC-Cy7 (clone 2D7/CCR5), anti-DC-SIGN v450 (clone DCN46) and anti-HLA-DR APC (clone L243) were from BD Biosciences (San Jose, CA, USA), anti-CD4 BV711 (clone OKT4) and anti-CD8 BV570 (clone RPA-T8) were from BioLegend (San Diego CA), anti-CD14 PE-Texas red was from Invitrogen, and anti-CD19 PE-Texas red (clone SJ25-C1) was from Abcam (Cambridge, UK). Data were acquired on a BD LSRFortessa instrument (BD Biosciences) and analyzed using FlowJo Version 9.7.5 software (TreeStar, Ashland, OR, USA). In some experiments, DCs were treated with 5 µg/ml Imiquimod (InvivoGen, Toulouse, France), 5 µg/ml Poly(I:C) (InvivoGen), 1 µg/ml ssRNA40LyoVec (Invivogen), 50 µM N-(n-Butyl)-deoxygalactojirimsin (NB-DGJ) (Toronto Research Chemicals, Toronto, Canada), 10 µM Chloroquine (InvivoGen), or with 5.6 µM TLR7 oligonucleotide antagonist IRS954: 5′-TGCTCCTGGAGGGGTTGT-3′ or the control oligo 5’-TCCTGCAGGTTAAGT-3 (Integrated DNA Technologies) (41 (link)).
Anti hla dr apc clone l243
Manufactured by BD
Sourced in United States
Anti-HLA-DR APC (clone L243) is a monoclonal antibody conjugated with the fluorescent dye Allophycocyanin (APC). It is designed to detect and quantify HLA-DR expression on the surface of cells.
Automatically generated - may contain errors
Lab products found in correlation
Lsr fortessa instrument, by BD (1 mentions)
Poly 1 c, by InvivoGen (1 mentions)
Ssrna40 lyovec, by InvivoGen (1 mentions)
Chloroquine, by InvivoGen (1 mentions)
Anti cd45 percp clone 2d1, by BD (1 mentions)
Imiquimod, by InvivoGen (1 mentions)
Imagestream x mark 2 imaging flow cytometer, by Merck Group (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Merck Group (1 mentions)
Cytofix cytoperm, by BD (1 mentions)
Perm wash buffer, by BD (1 mentions)
Facsverse, by BD (1 mentions)
Anti ki 67 pe clone b56, by BD (1 mentions)
3 protocols using anti hla dr apc clone l243
1
Flow Cytometric Analysis of Dendritic Cells
2
Flow Cytometric Analysis of B Cell Internalization
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Cells were stained with anti-HLA-DR APC (clone L243; BD Bioscience) in PBS supplemented with 0.1% BSA for 30 min in the dark on ice and fixed for 20 min in PBS with 4% PFA. Primary human B cells or Ramos B cells were washed and 4′,6′-diamidino-2-phenylindole (DAPI; Sigma) was added to stain the cell nucleus. Large particle internalization by human B cells was evaluated on an ImageStreamX mark II imaging flow cytometer (Merck). The acquired data was analyzed using IDEAS V6.2 Software (Merck) and FlowJo Software version 10 (Supplementary Figures 1A , 4 ).
3
Multiparameter Flow Cytometry of T Cells
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As described by Tian et al., fresh EDTA-K2 anticoagulated peripheral blood or frozen PBMCs were used for all the flow cytometry detection (Tian et al., 2015 (link)). The following anti-human flow cytometry antibodies cross-reactive with macaques were used: anti-CD3 APC—Cy7 (clone SP34–2), anti-CD8 PE-Cy7 (clone PRA-T8), anti-CD28 APC (clone CD28.8), anti-CD95 FITC (clone DX2) and anti-Ki67 PE (clone B56) were purchased from BD; anti-CD4 PerCP-Cy5.5 (clone OKT4), anti-CTLA-4 PE (clone BNI3), anti-HLA-DR APC (clone L243), and anti-PD-1 PE (clone EH12.2H7). They were purchased from Biolegend. Anti-CD38 FITC (Clone AT-1) was purchased from StemCell. CTLA-4 expression was detected by intracellular staining. Briefly, Cells were fixed and permeabilized with Cytofix/Cytoperm (BD BioSciences, USA) for 40 min at 4 °C in the dark after surface staining. Following two further washes with Perm/Wash buffer (BD BioSciences, USA), cells were intracellularly stained with anti-CTLA-4 for 1 h at 4 °C in the dark. Flow cytometric acquisitions were performed on a BD FACSVerse (BD BioSciences, USA) flow cytometer and all flow cytometric data were analyzed on FlowJoX10.0.7 software (FlowJo, USA).
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